Display of active beta-glucosidase on the surface of Schizosaccharomyces pombe cells using novel anchor proteins

作者：Tanaka Tsutomu; Matsumoto Sayoko; Yamada Mari; Yamada Ryosuke; Matsuda Fumio; Kondo Akihiko^*

来源：Applied Microbiology and Biotechnology, 2013, 97(10): 4343-4352.

DOI：10.1007/s00253-013-4733-0

• Schizosaccharomyces pombe
• Cell surface display
• Beta-glucosidase
• Cellobiose
• Anchor protein

Here, we demonstrate display of beta-glucosidase (BGL) on the surface of Schizosaccharomyces pombe cells using novel anchor proteins. A total of four candidate anchor proteins (SPBC21D10.06c, SPBC947.04, SPBC19C7.05, and SPBC359.04c) were selected from among almost all of S. pombe membrane proteins. The C-terminus of each anchor protein was genetically fused to the N-terminus of BGL, and the fusion protein was expressed using S. pombe as a host. The highest cell surface-associated BGL activity (107 U/10(5) cells was achieved with SPBC359.04c serving as the anchor, followed by SPBC947.04 (44 U/10(5) cells) and SPBC21D10.06c (38 U/10(5) cells). S. pombe displaying BGL with SPBC359.04c as an anchor showed the highest growth on 2 % cellobiose (10.7 x 10(7) cells/mL after 41 h of cultivation from an initial density of 0.1 x 10(7) cells/mL). Additionally, culturing BGL-displaying S. pombe in medium containing cellobiose as the sole carbon source did not affect protein expression, and ethanol fermentation from cellobiose was successfully demonstrated using BGL-displaying S. pombe. This is the first report describing a cell surface display system for the functionalization of S. pombe.

• 出版日期2013-5